{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T20:35:37Z","timestamp":1792787737951,"version":"4.5.0"},"reference-count":26,"publisher":"SAGE Publications","license":[{"start":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T00:00:00Z","timestamp":1791417600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"funder":[{"name":"National Centre for Disease Control-National One Health Programme for Prevention and Control of Zoonotic Diseases","award":["ANSCNIVEDICOL201900400103"],"award-info":[{"award-number":["ANSCNIVEDICOL201900400103"]}],"id":[{"id":"https:\/\/ror.org\/01ks7f963","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["Biopreservation and Biobanking"],"abstract":"<jats:sec>\n                    <jats:title>Introduction:<\/jats:title>\n                    <jats:p>\n                      A simple and reliable method for long-term preservation of\n                      <jats:italic toggle=\"yes\">Leptospira<\/jats:italic>\n                      spp. is essential for maintaining reference strains and field isolates for diagnostic and research purposes. Although 10% glycerol is commonly used as a cryoprotectant for\n                      <jats:italic toggle=\"yes\">Leptospira<\/jats:italic>\n                      preservation, the isolates in this study showed poor recovery, highlighting the need to optimize glycerol levels for cryopreservation in liquid nitrogen.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Objective:<\/jats:title>\n                    <jats:p>\n                      This study evaluated the storage of reference serovars\n                      <jats:italic toggle=\"yes\">Leptospira interrogans<\/jats:italic>\n                      serovar Australis (pathogenic),\n                      <jats:italic toggle=\"yes\">Leptospira fainei<\/jats:italic>\n                      serovar Hurstbridge (intermediate), and\n                      <jats:italic toggle=\"yes\">Leptospira biflexa<\/jats:italic>\n                      serovar Patoc (nonpathogenic) with 10% and 15% glycerol and 28 additional reference serovars, and 11 field isolates using 15% glycerol in liquid nitrogen.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods:<\/jats:title>\n                    <jats:p>Cultures were grown for 7 days in Ellinghausen\u2013McCullough\u2013Johnson\u2013Harris (EMJH) media supplemented with 10% rabbit serum and cryopreserved with 10% and 15% glycerol using a controlled-rate freezing protocol before storage at \u2212196\u00b0C. Viability was assessed monthly for up to 15 months using dark-field microscopy and Petroff\u2013Hausser chamber counts.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results:<\/jats:title>\n                    <jats:p>\n                      Both reference serovars (\n                      <jats:italic toggle=\"yes\">n<\/jats:italic>\n                      = 31) and field isolates (\n                      <jats:italic toggle=\"yes\">n<\/jats:italic>\n                      = 11) were successfully preserved using 15% glycerol and repeatedly recovered. Paired\n                      <jats:italic toggle=\"yes\">t<\/jats:italic>\n                      -test showed significant differences (\n                      <jats:italic toggle=\"yes\">p<\/jats:italic>\n                      &lt; 0.01) between 10% and 15% glycerol, while Kruskal\u2013Wallis analysis confirmed consistent cryoprotective efficiency of 15% glycerol across different\n                      <jats:italic toggle=\"yes\">Leptospira<\/jats:italic>\n                      species. Microscopic agglutination test confirmed unchanged antigenic properties, while hamster model studies demonstrated retained virulence, evidenced by acute leptospirosis.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion:<\/jats:title>\n                    <jats:p>\n                      This optimized protocol supports reliable long-term cryopreservation of\n                      <jats:italic toggle=\"yes\">Leptospira<\/jats:italic>\n                      spp. in liquid nitrogen without compromising viability, antigenicity, or virulence characteristics.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1177\/19475535261493709","type":"journal-article","created":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T14:44:29Z","timestamp":1791470669000},"update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":0,"title":["A Simple Modified Method for Long-Term Glycerol-Based Cryopreservation of\n                    <i>Leptospira<\/i>\n                    Spp. in Liquid Nitrogen"],"prefix":"10.1177","author":[{"given":"Malini","family":"Ponnidurai","sequence":"first","affiliation":[{"name":"ICAR-National Institute of Veterinary Epidemiology and Disease Informatics (NIVEDI), Bengaluru, India."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Archana","family":"Pal","sequence":"additional","affiliation":[{"name":"ICAR-National Institute of Veterinary Epidemiology and Disease Informatics (NIVEDI), Bengaluru, India."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vinod 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